Gasdermin D-mediated hepatocyte pyroptosis expands inflammatory responses that aggravate acute liver failure by upregulating monocyte chemotactic protein 1/CC chemokine receptor-2 to recruit macrophages

BACKGROUND Massive hepatocyte death is the core event in acute liver failure(ALF).Gasdermin D(GSDMD)-mediated pyroptosis is a type of highly inflammatory cell death. However, the role of hepatocyte pyroptosis and its mechanisms of expanding inflammatory responses in ALF are unclear.AIM To investigate the role and mechanisms of GSDMD-mediated hepatocyte pyroptosis through in vitro and in vivo experiments.METHODS The expression of pyroptosis pathway-associated proteins in liver tissues from ALF patients and a hepatocyte injury model was examined by Western blot.GSDMD short hairpin RNA(sh RNA) was used to investigate the effects of downregulation of GSDMD on monocyte chemotactic protein 1(MCP1) and its receptor CC chemokine receptor-2(CCR2) in vitro. For in vivo experiments, we used GSDMD knockout mice to investigate the role and mechanism of GSDMD in a D-galactose/lipopolysaccharide(D-Galn/LPS)-induced ALF mouse model.RESULTS The levels of pyroptosis pathway-associated proteins in liver tissue from ALFpatients and a hepatocyte injury model increased significantly. The level of GSDMD-N protein increased most obviously(P 0.001). In vitro, downregulation of GSDMD by sh RNA decreased the cell inhibition rate and the levels of MCP1/CCR2 proteins(P 0.01). In vivo, GSDMD knockout dramatically eliminated inflammatory damage in the liver and improved the survival of DGaln/LPS-induced ALF mice(P 0.001). Unlike the mechanism of immune cell pyroptosis that involves releasing interleukin(IL)-1β and IL-18, GSDMDmediated hepatocyte pyroptosis recruited macrophages via MCP1/CCR2 to aggravate hepatocyte death. However, this pathological process was inhibited after knocking down GSDMD.CONCLUSION GSDMD-mediated hepatocyte pyroptosis plays an important role in the pathogenesis of ALF, recruiting macrophages to release inflammatory mediators by upregulating MCP1/CCR2 and leading to expansion of the inflammatory responses. GSDMD knockout can reduce hepatocyte death and inflammatory responses, thus alleviating ALF.     

肾癌中调节性细胞死亡的新动向和未来展望

肾癌是泌尿系统常见的恶性肿瘤之一。近年来,我国肾癌的发病率呈逐年上升的趋势,严重威胁着人们的健康。调节性细胞死亡是由一种细胞主动有序的死亡方式,普遍存在于生命活动过程中,在维系生命活动的平衡中发挥着至关重要的作用。近期Science杂志上报道了一种新的调节性细胞死亡方式即铜死亡,进一步强化了生命体中细胞死亡的重要性。随着对调节性细胞死亡认识的不断深入,越来越多的研究显示不同的调节性细胞死亡（如铁死亡、焦亡、自噬等）均与肾癌的发生、发展密切相关。如诱导细胞铁死亡将显著抑制肾癌的侵袭和转移、并与肾癌患者的更好预后密切相关;细胞焦亡不仅可以诱导肾癌细胞死亡还可以激活抗肾癌的免疫应答;自噬在肾癌中具有“双向”作用,增强自噬可抑制肾癌细胞生长,但也可能减弱联合用药治疗的效果;抑制细胞凋亡和坏死性凋亡可以显著促进肾癌细胞的增殖、侵袭等。本文将综述铁死亡、细胞焦亡、自噬、细胞凋亡和坏死性凋亡的分子机制和在肾癌发生、发展中作用的研究进展并进行展望,为探索肾癌的发病机制和潜在的治疗靶点提供新的视角。     

基于数据库构建乳腺癌焦亡相关基因的预后风险模型

目的 探究焦亡相关差异表达基因(DEGs)在乳腺癌中预后价值并构建预后风险模型。方法 从癌症基因组图谱(TCGA)和肿瘤基因表达数据库(GEO)官网下载乳腺癌的基因测序、临床数据,筛选焦亡相关DEGs。将乳腺癌患者进行聚类分析。在TCGA队列中以最小绝对收缩和选择算子(LASSO)方法建立模型。利用Kaplan-Meier生存曲线、受试者工作特征曲线(ROC)、单因素及多因素Cox回归独立预后因素分析等评价该模型。GEO队列为验证集。通过GO、KEGG、ssGSEA分析风险DEGs的富集情况。结果 筛选出焦亡相关DEGs,聚类分析可见C2组总生存期(OS)延长,差异有统计学意义(P=0.020)。该模型K-M生存分析显示,高风险组OS缩短(TCGA队列中P<0.001,GEO队列中P=0.018)。ROC曲线下面积(AUC)表明该模型具有一定预测能力。单因素、多因素Cox回归分析表明,年龄,M、N分期和风险评分为OS的独立预测因子。GO、 KEGG富集与ssGSEA分析证实了风险相关DEGs与免疫炎症因子和通路有关。结论 研究构建了由9个焦亡相关基因组成的乳腺癌预后风险模型,为乳...     

IL-1β与IL-18在急性肺损伤中的表达及临床意义

目的 基于细胞焦亡理论研究白介素(IL)-1β与IL-18在急性肺损伤(ALI)中的表达及临床意义.方法 选取2016年1月至2017年1月在广州中医药大学第一附属医院重症医学科收治的符合ALI患者100例为观察组,期间同时招募50例健康体检者为对照组.酶联免疫法检测并比较两组受试者入组后IL-1β及IL-18的血清浓度水平,并比较观察组中存活患者与病死患者血清IL-1β与IL-18的含量.结果 观察组患者经常规治疗后,病死率为38.00%(38/100),而存活率为62.00%(62/100);入组后,两组受试者血清IL-18[(106.43±21.73)pg·mL-1 vs(174.29±36.31)pg·mL-1]与IL-1β[(62.14±15.88)pg·mL-1 vs(94.35±16.23)pg·mL-1]水平比较,对照组均明显低于观察组,差异均有统计学意义(P<0.05);在临床结局方面,病死组和存活组患者的IL-1β[(118.32±19.52)pg·mL-1 vs(87.86±16.49)pg·mL-1]与IL-18[(195.64±36.13)pg·mL-1 vs(136.74±23.09)pg·mL-1]水平比较,病死组均明显高于存活组,差异均有统计学意义(P<0.05).结论 细胞焦亡是ALI患者的重要病理机制之一,ALI的发生发展和IL-1β与IL-18的参与有关.     

Methods for monitoring cancer cell pyroptosis

Pyroptosis is a form of proinflammatory cell death that depends on the gasdermin family of proteins. The main features of pyroptosis are altered membrane permeability, cell swelling, membrane rupture, and the ability to mobilize a strong immune response. The relationship between pyroptosis and cancer has become a popular topic in immunological research. Multiple strategies for inducing pyroptosis in cancer cells have been developed for cancer therapy, including chemotherapy, small molecule drugs, and nanomedicines. In this review, we systematically discuss recent advances in research on the mechanisms of pyroptosis, and compare pyroptosis with apoptosis and necroptosis from several aspects. The development of various experimental systems has accompanied rapid progress in this field, but little consensus on monitoring pyroptosis is currently available. We focus on techniques commonly used to monitor pyroptosis, and describe future techniques that may be used to increase our knowledge in this field. Overall, the advancement of pyroptosis detection methods will help researchers to better investigate the relationships between pyroptosis and various cancers, and should provide insights into the use of these promising tools for cancer treatments.     

Shikonin from Chinese herbal medicine induces GSDME-controlled pyroptosis in tumor cellsOA

Objective:To investigate the potential anti-tumor mechanisms of naphthoquinone compound shikonin(SKN) extracted from the root of Chinese herbal medicine plant lithospermum(Lithospermum erythrorhizon Sieb. & Zucc.).Methods:We first observed that SKN treatment led to swelling and bubbles in HeLa cells that were similar to the phenotype of cell pyroptosis.Subsequently,the HeLa cells experienced a pyroptotic process with SKN,and this was then assessed using lactate dehydrogenase(LDH) release and...     

Eliciting pyroptosis to fuel cancer immunotherapy: mechanisms and strategies

Immune checkpoint blockade(ICB) therapy has recently shown promise in treating several malignancies. However, only a limited number of patients respond to this treatment, partially because of the “immune cold” condition of the tumor immune microenvironment. Pyroptosis is a type of gasdermin-mediated programmed cell death that often leads to inflammation and immune responses. Many studies on the mechanism and function of pyroptosis have led to increasing recognition of the role of pyroptosis in m...     

Non-apoptotic cell death-based cancer therapy:Molecular mechanism,pharmacological modulators,and nanomedicineOA

As an emerging cancer therapeutic target,non-apoptotic cell death such as ferroptosis,necroptosis and pyroptosis,etc.,has revealed significant potential in cancer treatment for bypassing apoptosis to enhance the undermined therapeutic efficacy triggered by apoptosis resistance.A variety of anticancer drugs,synthesized compounds and natural products have been proven recently to induce non-apoptotic cell death and exhibit excellent anti-tumor effects.Moreover,the convergence of nanotechnology with...     

益肾健脾泻浊中药对慢性肾脏病妊娠大鼠肾脏NLRP3/caspase-1/IL-1β信号通路及细胞焦亡的影响＊

目的 观察益肾健脾泻浊中药对慢性肾脏病妊娠大鼠肾脏NLRP3/caspase-1/IL-1β信号通路及细胞焦亡的影响。方法 将雌性Wistar大鼠随机分为6组，分别为对照组（Sham）、对照组 + 妊娠组（SP）、单侧输尿管结扎组（UUO）、UUO + 妊娠组（UP）、UUO + 妊娠 + 益肾健脾泻浊中药组（TCM）、UUO + 妊娠 + 依普利酮组（EPL）。UUO各组采用结扎单侧输尿管的方法复制慢性肾病模型，治疗组分别给予依普利酮100 mg?（kg?天）^-1和益肾健脾泻浊方3.11 g?（kg?天）^-1治疗。8周后妊娠各组大鼠按动情周期与雄鼠合笼，妊娠第19天处死母鼠。检测各组肾功能，醛固酮含量，采用SABC法及Western blot法检测核因子κB（Nuclear factor-kappa B，NF-κB）、核苷酸结合寡聚结构域样受体蛋白3 NOD［nucleotide-binding oligomerization domain-like receptor protein 3，NLRP3］、天冬氨酸特异性半胱氨酸蛋白1［Caspase-1］、白介素1β（Interleukine-1 beta，IL-1β）的表达，TUNEL法检测肾细胞DNA损伤情况。结果 与UUO组比较，UP组血清肌酐（Scr）、血尿素氮（BUN）和24 h尿蛋白及醛固酮含量较Sham组显著升高（P < 0.05），治疗后，两给药组Scr、BUN、24 h尿蛋白、醛固酮显著降低（P < 0.05）。与Sham组比较，SP组大鼠肾组织NF-κB、NLRP3、Caspase-1、IL-1β表达明显升高（P < 0.05），TUNEL阳性细胞少量增加（P < 0.05）。NLRP3炎症小体主要表达于浸润巨噬细胞及肾小管上皮细胞；Caspase-1和IL-1β主要表达于肾小管上皮细胞胞浆；TUNEL阳性细胞主要见于远端小管上皮细胞。与UUO组比较，UP组大鼠肾组织NF-κB、NLRP3、Pro-caspase-1、Caspase-1、Pro-IL-1β、IL-1β指标表达也明显升高（P < 0.05），TUNEL阳性细胞明显增多（P < 0.05）。与UP组比较，EPL、TCM组大鼠肾组织上述指标表达明显降低（P < 0.05），TUNEL阳性细胞明显减少（P < 0.05）。两个给药组间各项指标无明显差异。结论 益肾健脾泻浊方及依普利酮可缓解慢性肾病妊娠大鼠肾脏炎症损伤，下调肾脏NLRP3炎症小体信号通路分子表达，从而抑制细胞焦亡，减轻肾脏炎症损伤。